• 제목/요약/키워드: differential gel electrophoresis

검색결과 65건 처리시간 0.032초

Studies on the Processing Properties and Interactions Between Porcine Blood Proteins and Waxy Rice Starch During Making Porcine Blood Cake

  • Lin, Chin-Wen;Yang, Jeng-Huh;Chu, Hsien-Pin;Su, Ho-Ping;Chen, Hsiao-Ling;Huang, Chia-Cheong
    • Asian-Australasian Journal of Animal Sciences
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    • 제14권3호
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    • pp.358-364
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    • 2001
  • The physiochemical properties and interactions between porcine blood and waxy rice were determined. Addition of calcium chloride (0.15%) improved acceptability of blood cake and increased the gelatinization degree of waxy rice. The water-holding capacity of porcine blood gel (blood/water=60/40, v/v), extent of absorption and gelatinization of waxy rice, and scanning electron microscopy showed that blood protein matrix and waxy rice are competitors for holding water in the cooking procedure. Non-haem iron content increased linearly (R=0.95) when heating temperature rose. The presence of blood proteins caused increasing of peak temperature (Tp) of gelatinization in differential scanning calorimetric thermal gram, The microstnlcture of plasma proteins and haemoglobin appeared continuous changes, and interacted with surface of waxy rice flour in terms of network and mosaic form, respectively. The electrophoretic patterns revealed an interaction between plasma proteins and waxy rice glutelin and haemoglobin when heated could be found at temperatures above $60^{\circ}C$.

Differential Role of protein Kinase C in Ginsenoside $Rh_2$ - induced Apoptosis in SK-N-BE(2) and C6Bu-1 Cells

  • Young Sook Kim;Sun
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 1998년도 Advances in Ginseng Research - Proceedings of the 7th International Symposium on Ginseng -
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    • pp.244-252
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    • 1998
  • Ginsenoside Rh, (G-Rh,) from Panax ginseng induced morphological features of apoptosis and DNA fragmentation as a biochemical marker of apoptosis confirmed by TUNEL reaction and agarose gel electrophoresis in human neuroblastoma SK-N-BE(2) and rat glioma C6Bu-1 cells During apoptosis by G-Rh2, protein kinase C (PKC) isoforms were analysed by immunoblotting. In SK-N-BE(2) cells, the levels of a, p and ${\gamma}$ subtypes were increased by undergoing apoptosis, while PKC e isoform increased early in treatment (3 h and 6 h). In addition, PKC s isoform gradually decreased during apoptosis by G-Rh2 and PKC $\theta$ isoform was detected in neither untreated- nor G-Rh1-treated SK-N-BE(2) cells (data not shown). However, no significant changes in the level of S and s isoforms were observed in C6Bu-1 cells undergoing apoptosis by G-Rh2. These results suggest that PKC subtypes may play differential roles in apoptotic signal pathways and their roles can be cell type-specific in apoptosis induced by G-Rh2.

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Proteomic Approach Analysis of Mammary Membrane Proteins Expression Profiles in Holstein Cows

  • Yang, Yong-xin;Cao, Sui-zhong;Zhang, Yong;Zhao, Xing-xu
    • Asian-Australasian Journal of Animal Sciences
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    • 제22권6호
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    • pp.885-892
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    • 2009
  • To investigate host defense mechanisms for protecting the mammary gland from mastitis infection, the membrane fraction of mammary tissues from Holstein cows was purified by differential velocity centrifugation, and then the sodium dodecyl sulfate-polyacrylamid gel electrophoresis (SDS-PAGE) separated proteins were identified by ion trap mass spectrometer equipped with a Surveyor high performance liquid chromatography (HPLC) system. A total of 183 proteins were identified. Bioinformatics software was applied to analyse physicochemical characteristics of the identified proteins and to predict biochemical function. These data may provide valuable information to investigate the mechanisms of mammary gland milk secretion and infectious disease, and enable a clear identification of proteins and potential protein targets for therapies.

Proteomics-driven Identification of Putative AfsR2-target Proteins Stimulating Antibiotic Biosynthesis in Streptomyces lividans

  • Kim Chang-Young;Park Hyun-Joo;Kim Eung-Soo
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제10권3호
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    • pp.248-253
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    • 2005
  • AfsR2, originally identified from Streptomyces lividans, is a global regulatory protein which stimulates antibiotic biosynthesis. Through its stable chromosomal integration, the high level of gene expression of afsR2 significantly induced antibiotic production as well as the sporulation of S. lividans, implying the presence of yet-uncharacterized AfsR2-target proteins. To identify and evaluate the putative AfsR2-target proteins involved in antibiotic regulation, the proteomics-driven approach was applied to the wild-type S. lividans and the afsR2-integrated actinorhodin overproducing strain. The 20 gel-electrophoresis gave approximately 340 protein spots showing different protein expression patterns between these two S. lividans strains. Further MALDI-TOF analysis revealed several AfsR2-target proteins, including glyceraldehyde-3-phosphate dehydrogenase, putative phosphate transport system regulator, guanosine penta phosphate synthetase/polyribonucleotide nucleotidyltransferase, and superoxide dismutase, which suggests that the AfsR2 should be a pleiotropic regulatory protein which controls differential expressions of various kinds of genes in Streptomyces species.

Proteome Analysis of Vernalization-Treated Arabidopsis thaliana by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

  • Cho, Mi-Ran;Lee, Kyung-Hyeon;Hyun, You-Bong;Lee, Il-Ha;Kim, Hie-Joon
    • Bulletin of the Korean Chemical Society
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    • 제28권3호
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    • pp.427-431
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    • 2007
  • In order to gain insight into the molecular changes at the protein level in plants exposed to low temperature for a long period of time (vernalization), proteome analyses of vernalization-treated Arabidopsis thaliana have been carried out by two-dimensional gel electrophoresis followed by matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry. Fourteen proteins including ATP binding/GTP binding/translation elongation factor and glycine-rich RNA-binding protein 7 (GRP7) showed differential expression in vernalization-treated Arabidopsis thaliana. GRP7 showed the most dramatic increase in expression suggesting its involvement in response to vernalization treatment.

Proteomic Analysis of Drought Stress-Responsive Proteins in Rice Endosperm Affecting Grain Quality

  • Mushtaq, Roohi;Katiyar, Sanjay;Bennett, John
    • Journal of Crop Science and Biotechnology
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    • 제11권4호
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    • pp.227-232
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    • 2008
  • Drought stress is one of the major abiotic stresses in agriculture worldwide. We report here a proteomic approach to investigate the impact of post-fertilization drought on grain quality in rice seed endosperm (Oryza sativa cv. IR-64). Plants were stressed for 4 days at 3 days before heading. Total proteins of endosperm were extracted and separated by two-dimensional gel electrophoresis. Not many protein spots showed differential accumulation in drought-stressed samples. More than 400 protein spots were reproducibly detected, including three that were up-regulated and five down-regulated. Mass spectrometry analysis and database searching helped us to identify six spots representing different proteins. Functionally, the identified proteins were related to protein synthesis and carbohydrate metabolism, such as Granule-Bound Starch Synthase (GBSS, Wx protein), which is thought to play a very important role in starch biosynthesis and quality, a very crucial factor in determining rice grain quality.

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New Protein Extraction/Solubilization Protocol for Gel-based Proteomics of Rat (Female) Whole Brain and Brain Regions

  • Hirano, Misato;Rakwal, Randeep;Shibato, Junko;Agrawal, Ganesh Kumar;Jwa, Nam-Soo;Iwahashi, Hitoshi;Masuo, Yoshinori
    • Molecules and Cells
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    • 제22권1호
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    • pp.119-125
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    • 2006
  • The rat is an accepted model for studying human psychiatric/neurological disorders. We provide a protocol for total soluble protein extraction using trichloroacetic acid/acetone (TCA/A) from rat (female) whole brain, 10 brain regions and the pituitary gland, and show that two-dimensional gel electrophoresis (2-DGE) using precast immobilized pH (4-7) gradient (IPG) strip gels (13 cm) in the first dimension yields clean silver nitrate stained protein profiles. Though TCA/A precipitation may not be "ideal", the important choice here is the selection of an appropriate lysis buffer (LB) for solubilizing precipitated proteins. Our results reveal enrichment of protein spots by use of individual brain regions rather than whole brain, as well as the presence of differentially expressed spots in their proteomes. Thus individual brain regions provide improved protein coverage and are better suited for differential protein detection. Moreover, using a phosphoprotein-specific dye, ingel detection of phosphoproteins was demonstrated. Representative high-resolution silver nitrate stained proteome profiles of rat whole brain total soluble protein are presented. Shortcomings apart (failure to separate membrane proteins), gel-based proteomics remains a viable option, and 2-DGE is the method of choice for generating high-resolution proteome maps of rat brain and brain regions.

검정곰팡이의 분화(分化)에 따르는 균체내(菌體內)의 고인산(高燐酸)뉴크레오티드의 소장(消長)에 관한 연구(硏究) (Studies on the Highly-phosphorylated Nucleotides during the Differentiation of Aspergillus niger)

  • 김종협
    • 한국균학회지
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    • 제10권2호
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    • pp.57-65
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    • 1982
  • 경정곰팡이(Aspergillus niger)의 포자형성(胞子形成)을 액침배양법(液浸培養法)과 동조적방법(同調的方法)으로 여행(旅行)하면서 인산화합물(燐酸化合物), 고인산화(高燐酸化)뉴크레오티드, 및 RNA(핵산)(核酸)의 동태(動態)를 연구(硏究)하였다. 결과(結果)는 다음과 같이 요약(要約)된다. 1. 곰팡이의 포자형성시(胞子形成時)에 유기인산화합물(有機燐酸化合物)의 양(量)이 증가(增加)하였으며, 무기(無機) Polyphosphate의 양(量)은 반대(反對)로 감소(減少)하였다. 2. 무기인산양(無機燐酸量)은 변동(變動)하지 않았으며, 총인산화합물(總燐酸化合物)의 양(量)은 초기균사생장시(初期菌絲生長時)에는 감소(減少)하고 포자낭병시기(胞子囊柄時期)부터는 변동(變動)하지 않았다. 3. RNA핵산구획(核酸區劃)에 있어서 orcinol 반응양성구(反應陽性區)의 흡광도(吸光度)가 급격(急激)히 증가(增加)하였다. 그러나 UV흡광도(吸光度)는 완만하게 증가(增加)하였다. 4. 포자낭병(胞子囊柄)및 포자형성시기(胞子形成時期)에 guanosinetetraphosphate가 검출현(檢出現) 되었다. 5. 고분자양성(高分子量性) RNA (rRNA분획(分劃))의 polyacrylamide gel 전기영동(電氣泳動)을 2.6% gel로서 시행(施行)한 바 polyphosphate와 RNA의 결합물(結合物)이 상당양(相當量) 존재(存在)함을 알았다. 이상(以上)의 결과(結果)로 보아 검정곰팡이의 분화과정(分化過程)에 있어서 고인산화(高燐酸化) RNA 화합물(化合物)의 존재(存在)가 확인(確認)되었으며, 이 화합물(化合物) 가운데 guanosine tetraphosphate (G4P)의 존재(存在)가 진핵미생물(眞核微生物)인 곰팡이에서도 검출(檢出)되었음을 지적(指摘)한다.

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Differential Proteome Analysis of Breast and Thigh Muscles between Korean Native Chickens and Commercial Broilers

  • Liu, Xian De;Jayasena, Dinesh D.;Jung, Yeon-Kuk;Jung, Samooel;Kang, Bo-Seok;Heo, Kang-Nyeong;Lee, Jun-Heon;Jo, Cheo-Run
    • Asian-Australasian Journal of Animal Sciences
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    • 제25권6호
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    • pp.895-902
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    • 2012
  • The Korean native chickens (Woorimotdak$^{TM}$, KNC) and commercial broilers (Ross, CB) show obvious differences in meat flavor after cooking. To understand the contribution of protein and peptide for meat flavor, 2-dimensional (2-D) gel electrophoresis and matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry was performed. A total of 16 protein spots were differentially expressed in the breast and thigh meat between the two breeds. A total of seven protein spots were represented by different levels between KNC and CB for breast meat. Among them three protein spots (TU39149, TU40162 and TU39598) showed increases in their expressions in KNC while other four protein spots (BU40125, BU40119, BU40029 and BU39904) showed increases in CB. All nine protein spots that were represented by different levels between KNC and CB for thigh meat showed increases in their expression in KNC. Phosphoglucomutase 1 (PGM 1), myosin heavy chain (MyHC), heat shock protein B1 (HSP27), cytochrome c reductase (Enzyme Q), Glyoxylase 1, DNA methyltransferase 3B (DNA MTase 3) were identified as the main protein spots by MALDI-TOF mass spectrometry. These results can provide valuable basic information for understanding the molecular mechanism responsible for breed specific differences in meat quality, especially the meat flavour.

LPS로 자극된 macrophage RAW264.7 세포에서 ascochlorin에 대한 단백질체 분석 (Proteome Analysis of Responses to Ascochlorin in LPS-induced Mouse Macrophage RAW264.7 Cells by 2-D Gel Electrophoresis and MALDI-TOF MS.)

  • 장영채
    • 생명과학회지
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    • 제18권6호
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    • pp.814-825
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    • 2008
  • 아스코크로린(Ascochlorin, ASC)은 Ascochyta viciae로부터 추출된 프레닐페놀 물질로, 혈청 콜레스테롤과 트리글리세라이드 수치를 감소시키고 종양 성장을 억제한다는 연구 결과가 보고되어 있다. 본 논문에서는 아스코크로린이 생리학적 혹은 병리학적인 작용과 염증반응에서 약리학적으로 유도되는 반응을 어떻게 조절하며, 이러한 메커니즘에 대해 이해하기 위해 mouse macrophage Raw264.7 세포에 아스코크로린을 처리하여 이에 대한 프로테옴의 특이적인 발현에 대해 분석하였다. 따라서 본 연구는 LPS를 처리한 mouse macrophage Raw264.7 세포에 아스코크로린을 처리하여 염증과정에 관련된 단백질의 발현 양상을 확인하기 위해 프로테오믹스를 시행하였다. Mouse macrophage RAW264.7 세포에 아스코크로린을 처리한 조건과 무처리한 조건으로 나누어 two-dimensional electrophoresis (2-D SDS-PAGE), matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF-MS)와 bioinformatics 방법으로 아스코크로린을 처리한 mouse macrophage Raw264.6 세포의 프로테옴을 분석하였다. 그 결과 mouse macrophage Raw264.7 세포에 아스코크로린 처리 시 Calreticulin이 4배 감소, ${\beta}-actin$도 4배 감소 그리고 vimentin이 1.5배 감소함을 확인 할 수 있었다. 그러나 rabaptin 아스코크로린 처리에 의해 3배 증가함을 확인 할 수 있었다. 이러한 단백질 발현은 RT-PCR을 수행하여 결과에 대해 재확인 하였으며, 프로테오믹스와 동일한 결과를 얻을 수 있었다. 따라서 본 연구를 통해 LPS 처리에 의해 활성화된 mouse macrophage RAW264.7 세포에 ASC를 처리한 후 이차원 전기영동법을 이용하여, 단백질의 발현 변화 및 양상을 규명하고 단백질 지도를 확립 하였으며, RAW264.7 세포를 이용한 면역세포 모델에서 ASC의 항염증 작용을 중심으로 생리활성 조절기능을 확인 할 수 있었다. 향후 분자 기능 조절 연구와 전 임상 연구를 통해 ASC의 생리활성 조절 기능을 규명한다면 ASC는 항염증 및 항암활성을 갖는 약물로 개발될 것으로 기대된다.